1. Field of the Invention
Embodiments of the invention described herein relate to the field of nanotechnology and are more specifically directed to a microfluidic multi-compartment device for neuroscience research.
2. Background
A multi-compartment culturing device for neuritic isolation was first described by Campenot for primary cultures of sympathetic neurons. In this method, a tissue culture dish is coated with collagen and parallel lines, spaced 200 um apart, are scratched along the surface of the dish. A three-compartment Teflon piece is sealed to a Petri dish with silicone grease and neurons are plated in the small central chamber of the Teflon piece. Nuerites grow outwards into the two other compartments on either side, aligning parallel to the scratches. Variations of the Campenot chamber have been used in studies of various types of long projection neurons. However the Campenot chamber did not work well when used to culture cortical and hippocampal neurons.
Ivins, et al. developed a chamber designed for cortical and hippocampal neuron cultures using a relatively short barrier distance (150 um versus 300 um in the classic Campenot chamber). These chambers use a glass coverslip fixed to hemisected Teflon tubing using Sylgard 184 (Dow Corning, Corning N.Y.). A small amount of silicone vacuum grease is applied to the bottom of the converslip using a dissecting microscope and the whole apparatus is placed on the tissue culture dish. Neurites extend through the vacuum grease barrier between the polystyrene and the coverslip, if the vacuum grease barrier is sufficiently thin. A problem with these devices is that the process of making the chambers is laborious and their successfulness is directly related to the skill level of the individual using the device. Additionally, there is no alignment of neurons and the apparatus is not compatible with live cell imaging, thus, the effects of insults were observed only after the cells were fixed.